The invention relates to acyclic sulfamides and bis-sulfamides useful as inhibitors of proteolytic enzymes. The invention further relates to general templates for use in designing combinatorial libraries, screening and identification of biologically active, peptidomimetic protease inhibitors. There are four classes of proteases: serine, cysteine, aspartic and metalloproteases.
Degenerative diseases associated with serine proteases such as human leukocyte elastase (HLE) include cystic fibrosis, chronic obstructive pulmonary disease (e.g., emphysema and asthma), adult respiratory distress syndrome (ARDS), inflammatory bowel disease, chronic bronchitis, psoriasis, rheumatoid arthritis, pancreatitis, periodontal disease, and other inflammatory diseases
A variety of cyclic mechanism-based inhibitors of serine proteases are known. Exemplary protease inhibitors include sulfonyloxy derivatives of isothiazolidin-3-one 1,1-dioxides (Groutas et al., Biochemical and Biophysical Research Communications 1997(2):730 (December, 1993)), 1,2,5-thiadiazolidin-3-one 1,1-dioxides (Kuang et al., J. Am. Chem. Soc. 121:8128 (September 1999)) and 3-alkyl-N-hydroxysuccinimides (Groutas et al., J. Med. Chem. 32:1607 (1989)). These compounds react irreversibly with a variety of serine proteases via a sequence of steps involving binding of the inhibitor molecule to the active site of the enzyme, nucleophilic ring opening of the cyclic inhibitor, rearrangement of the ring-opened structure to provide a reactive intermediate, followed by irreversible reaction of the compound with a second site on the enzyme, resulting in enzyme deactivation.
Diseases associated with cysteine proteases include cancer metastasis, osteoporosis and osteoarthritis (McGrath et al. Nature: Structural Biology 4(2):105 (1997)), bone resorption, muscular dystrophy, parasitic diseases (leishmaniasis, malaria) (Li et al. Bioorg. Med. Chem. 4(9):1421 (1996); Rosenthal et al. J. Clin. Invest. 91:1052 (1993)), inflammation, common cold (Webber et al. J. Med. Chem. 39:5072 (1996)), and hepatitis A (Malcolm et al. Biochemistry 34:8172 (1996)). Many known cysteine protease inhibitors are peptidyl aldehydes, halomethylketones and Michael acceptors (xcex1,xcex2-unsaturated groups).
Matrix metalloproteinases (MMPs), such as collagenase, stromelysin and gelatinase, are involved in connective tissue breakdown. Metalloproteinase (MMP) inhibitors are of potential value in the treatment of neuroinflammatory disorders, including those involving myelin degradation, for example, multiple sclerosis, as well as management of angiogenesis-dependent diseases, proliferative retinopathies, neovascular glaucoma, ocular tumors, angiofibromas and hemangiomas. See, WO 95/35275, entitled xe2x80x9cMetalloproteinase Inhibitorsxe2x80x9d. Many known metalloproteinases are characterized by the presence in the structure of a zinc(II) ion at the active site, and thus, most known MMP inhibitors typically include hydroxamic acid or carboxylic acid to bind zinc. For example, arylsulfonamide-substituted hydroxamic acids have been reported as matrix metalloproteinase inhibitors. See, U.S. Pat. No. 5,455,258 to MacPherson et al.
The present invention makes available a new class of compounds useful as protease inhibitors. The open sulfamide structure is anticipated to be more stable chemically than related closed ring structures, e.g., 1,2,5-thiadiazolidin-3-one 1,1-dioxides, yet show high inhibition of proteolytic activity.
The invention features a compound having the general formula I 
and pharmaceutically acceptable salts thereof, wherein,
Z is a chemical species or R1 capable of binding at a primary specificity site of a protease;
Y is a chemical species reactive to a specific class of protease;
each of R2, R3, R5 and R7 is independently selected from the group consisting hydrogen, alkyls, aryls, substituted aryls, alkylaryls and arylalkyls;
R4 and R6 are independently selected from the group consisting of:
(a) H, alkyl, aryl, arylalkyl, alkylaryl, substituted derivatives thereof, and Ri;
(b) xe2x80x94C(O)OH and derivatives thereof, said derivatives selected from the group consisting of xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94C(O)[NHCHRi(q)C(O)]qNRYRZ; and
(c) xe2x80x94CHRiNH2 and derivatives thereof, said derivatives selected from the group consisting of xe2x80x94CHRiNHW, xe2x80x94CHRiNHC(O)OQ, xe2x80x94CHRiNHC(O)R, xe2x80x94CHRiNHC(O)NRYRZ, xe2x80x94CHRiNHC(O)[NHCHRi(q)C(O)]qOQ, xe2x80x94CHRiNHSO2R, and xe2x80x94CHRiNH[C(O)CHRi(r)NH]rW,
where q and r independently are integers from 1 to 10 inclusive;
where J is a carboxyl protecting group;
where G is an amino protecting group;
where Q is H, R or J;
where W is H, R or G;
where each Ri is independently selected from naturally or non-naturally occurring amino acid side chains;
where R is alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical; and
where each RY and RZ is independently H, alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical.
In another aspect of the invention, a compound of the general formula III, is provided, and pharmaceutically acceptable salts thereof, 
wherein
Z is a chemical species or Ri capable of binding at a primary specificity site of a protease;
Y is a chemical species reactive to a specific class of protease;
R30 is selected from the group consisting of hydrogen, alkyls, aryls, substituted aryls, alkylaryls or arylalkyls;
R31 is selected from the group consisting of alkyl, aryl, arylalkyl, alkylaryl, substituted derivatives thereof, Ri, xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)R, xe2x80x94SO2R xe2x80x94[C(O)CHRi(r)NH]rW, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94C(O)[NHCHRi(q)C(O)]qNRYRZ; and
R32 is selected from the group consisting of alkyl, aryl, substituted aryl, arylalkyl, alkylaryl, Ri; xe2x80x94CHRiC(O)O, xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94[C(O)CHRi(r)NH]rW;
where q and r independently are integers from 1 to 10 inclusive,
where Q is H, R or J, and J is a carboxyl protecting group,
where W is H, R or GI and G is an amino protecting group;
where each Ri is independently selected from naturally or non-naturally occurring amino acid side chains,
where R is alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical, and
where each RY and RZ is independently H, alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical.
In another aspect of the invention, a compound of the general formula IV is provided, and pharmaceutically acceptable salts thereof, 
wherein
Z is a chemical species or Ri capable of binding at a primary specificity site of a protease;
Y is a chemical species reactive to a specific class of protease;
R30 is selected from the group consisting of hydrogen, alkyls, aryls, alkylaryls or arylalkyls;
R34 is aryloxy, arylalkyl, xe2x80x94CHRiNHW or xe2x80x94NHCHRiC(O)OQ; and
R35 is alkyl, aryl, alkylaryl, arylalkyl or amino acid side group, Ri,
where Q is H, R or a carboxyl protecting group;
where W is H, R or an amino protecting group; and
where R is H, alkyl, aryl, substituted aryl, alkaryl, aralkyl, or heterocyclic radical.
In another aspect of the invention, a compound of the general formula V is provided and pharmaceutically acceptable salts thereof, 
wherein
Z is a chemical species or Ri capable of binding at a primary specificity site of a protease;
Y is a chemical species reactive to a specific class of protease;
R30 is selected from the group consisting of hydrogen, alkyls, aryls, alkylaryls or arylalkyls;
R36 is alkyl, aryl, alkylaryl, arylalkyl or amino acid side group, Ri.;
R34 is aryloxy, arylalkyl, xe2x80x94CHRiNHW or xe2x80x94NHCHRiC(O)OQ,
where Q is H, R or a carboxyl protecting group;
where W is H, R or an amino protecting group; and
where R is H, alkyl, aryl, substituted aryl, alkaryl, aralkyl, or heterocyclic radical.
In still another aspect of the invention, a compound having the general formula II, and pharmaceutically acceptable salts thereof, is provided, 
wherein
Z is a chemical species or Ri capable of binding at a primary specificity site of a protease inhibitor;
Y is a chemical species reactive to a specific class of protease inhibitor;
each of R2, R3, R5, R7 and R8 is independently selected from the group consisting of hydrogen, and saturated, unsaturated and aromatic hydrocarbons, and more particularly, hydrogen, alkyls, aryls, alkylaryls or arylalkyls; and
R4 and R6 are independently selected from the group consisting of:
(a) H, alkyl, aryl, arylalkyl, alkylaryl, substituted derivatives thereof, and Ri;
(b) xe2x80x94C(O)OH and derivatives thereof, said derivatives selected from the group consisting of xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94C(O)[NHCHRi(q)C(O)]qNRYRZ; and
(c) xe2x80x94CHRiNH2 and derivatives thereof, said derivatives selected from the group consisting of xe2x80x94CHRiNHW, xe2x80x94CHRiNHC(O)OQ, xe2x80x94CHRiNHC(O)R, xe2x80x94CHRiNHC(O)NRYRZ, xe2x80x94CHRiNHC(O)[NHCHRi(q)C(O)]qOQ, xe2x80x94CHRiNHSO2R, and xe2x80x94CHRiNH[C(O)CHRi(r)NH]rW,
where q and r independently are integers from 1 to 10 inclusive;
where J is a carboxyl protecting group;
where G is an amino protecting group;
where Q is H, R or J;
where W is H, R or G;
where each Ri is independently selected from naturally or non-naturally occurring amino acid side chains;
where R is alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical; and
where each RY and RZ is independently H, alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical.
In yet another aspect of the invention, a compound of the general formula VI, and pharmaceutically acceptable salts thereof, is provided, 
wherein
Z is a chemical species or Ri capable of binding at a primary specificity site of a protease;
Y is a chemical species reactive to a specific class of protease;
R30 is selected from the group consisting of hydrogen, alkyls, aryls, alkylaryls or arylalkyls;
R31 is selected from the group consisting of alkyl, aryl, arylalkyl, alkylaryl, substituted derivatives thereof, Ri, xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)R, xe2x80x94SO2R xe2x80x94[C(O)CHRi(r)NH]rW, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94C(O)[NHCHRi(q)C(O)]qNRYRZ; and
R32 is selected from the group consisting of alkyl, aryl, substituted aryl, arylalkyl, alkylaryl, Ri; xe2x80x94CHRiC(O)O, xe2x80x94C(O)OQ, xe2x80x94C(O)NRYRZ, xe2x80x94C(O)[NHCHRi(q)C(O)]qOQ, and xe2x80x94[C(O)CHRi(r)NH]rW;
where q and r independently are integers from 1 to 10 inclusive,
where Q is H, R or J, and J is a carboxyl protecting group,
where W is H, R or G, and G is an amino protecting group;
where each Ri is independently selected from naturally or non-naturally occurring amino acid side chains,
where R is alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical, and
where each RY and RZ is independently H, alkyl, aryl, substituted aryl, alkylaryl, arylalkyl, or heterocyclic radical.
The invention features a universal template for creating biologically active, peptidomimetic compounds, such as those of formulae I-VI, above. The chemical stability, side chain orientation, and polarity characteristics of the disclosed core template combine to provide numerous inhibitors of a variety of enzymes, including serine and cysteine proteases and matrix metalloproteases (MMPs) and aspartic proteases and others. The disclosed compounds are designed to have inhibitory activity, including selectivity and improved subsite interactions, by varying the pendant groups of the template compounds.
The disclosed inhibitors are useful in methods of treating a protease-related condition, such as a degenerative disease, wherein a pharmaceutically effective amount of a composition including one or more disclosed inhibitors is administered to a patient. The invention therefore further includes methods of reducing or inhibiting protease activity by contacting a protease with one of the compounds I-VI above. The protease may be from any protease source, including human and non-human mammals, and tissues, cells or membranes isolated therefrom which include the protease, or an isolated enzyme which has a binding affinity for the compounds of the invention. Binding affinity is defined as having a Ki on the order of at least micromolar (xcexcM) and preferably nanomolar (nM).
Other features and advantages of the invention will be apparent from the detailed description and examples below.